IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS)

e-ISSN: 2319-2380, p-ISSN: 2319-2372. Volume 8, Issue 4 Ver. II (Apr. 2015), PP 45-50
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Quality Assessment Of Unbranded Refined Palmkernel Oil In

Distribution Within Five Local Markets In Port Harcourt, Rivers
State Nigeria
Ezediokpu Marycolette .N1, Omorodion, Nnenna .J.P2, Okerentugba P.O3,
and Ego C.C4.
Department of Microbiology, University of Port Harcourt, Choba,
P.M.B 5323 Port Harcourt, River State, Nigeria.

Abstract: The study was conducted to evaluate the quality of groundnut oil sold in the five major markets in
Port Harcourt River state, Nigeria. Samples were collected from Rumuokoro market, Choba market, Mile 1
market, Mile 3 market and Oil mill market. Microbial quality such as; Total heterotrophic bacterial
load(THBC),Total Heterotrophic Fungal count(THFC) Total lipid utilizing bacterial and fungal counts (TLUB
and (TLUF) , Total coliform count as ,well as physicochemical qualities such as; iodine number, relative
density, pH, peroxide value and free fatty acid(ffa) content were investigated by standard methods. The results
obtained shows that the total heterotrophic bacterial load ranged from 1.2107 - 1.51010. Total heterotrophic
fungal count ranged from 3.3 105 -1.64 106, lipid utilizing bacteria (LUB) ranges from 6.0106 -1.4109
while the lipid utilizing fungi ranges from 3.010510 7.0106 . Bacteria isolates tentatively identified and
their frequency of occurrence in all the samples were Staphylococcus sp. (100%)., Alcaligenes sp.(60%).,
Enterococcus sp. (100%)., Salmonella sp. (100%)., Achromobacter sp. (40%)., Micrococcus sp. (40%)., and
Klebsiella sp. (20%). Fungal isolates identified as Aspergillus sp., Penicillium sp. and Fusarium sp. Results
obtained from the physicochemical analysis showed that Iodine number(wijis) was between 17.80-15.86,
Relative density(g) 0.94-0.905, pH 5.3, Peroxide value(Meq/kg) 59.92-59.48 and free fatty acid (%) 1.57-1.29.
Results on the microbiological and physicochemical qualities were found to exceed acceptable limits, this
suggests that palm kernel oil in distribution within Port Harcourt metropolis are of low quality and requires
some improvement in the production process.
Keywords : Groundnut oil, Quality, Microbial, Physicochemical.

I.

Introduction

Palm kernel oil is obtained from palm kernel seeds and grows widely in certain locations such as Niger
Delta and South Eastern parts of Nigeria (Komolafe and Adegbola, 1980; Samuel and Alabi, 2012). The palm
kernel oil is extracted from the walnut which has a hard hill or shell called kernel. It is also called lauric oil
because it has high content of lauric acid (Deutheux, 2004;Macaire et al., 2010) the extraction of palm kernel oil
is made by various techniques and the obtained oil is used in the food domain as well as non-domains. In the
food domain, it is used in the preparation of certain traditional dishes and enter also in the constitution of food
fats. In the non-food domain, its higher proportion of lauric acid gives to the oil an important characteristics
used in the industries of beauty care and soap factory. This property also characterizes, its use in traditional
pharmacopoeia ( Salmiah et al., 1998; Macaire 2010). The acceptability of the products at world edible oil
market depends on its ability to satisfy basic standard tests for fats and oil (Takakura, 2002; Samuel and Alabi,
2012).

II.

Materials And Methods

2.1 Collection of Sample:

Sampling were carried out using the modified method by Chabiri et al., ( 2009).Palm kernel oil
(Elaeis guinensis) is derived from palm kernel mills. Samples were similarly collected from different local
sellers within the same market in Port Harcourt. Five local markets were selected, these include; Rumuokoro
market, Oil mill, Mile one (1), Mile three (3) and Choba markest. Fifty mililitres of palm oil are collected from
10 different spots from different local sellers to make a composite of 500ml collected in a well labeled sterile
bottle. For each market the sum total of 50 sample were composited into 5 samples, each per market.
2.2 Maintenance of Sample:
Samples were maintained at ambient temperature in sterile containers and were quickly transported in
sterile canisters to the laboratory for analysis.
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Quality Assessment Of Unbranded Refined Palmkernel Oil In Distribution Within Five Local
2.3 Preparation of Sample:
Samples were placed in sterile labeled bottle and were first diluted in Ringers solutions(0.12g/l calcium
carbonate ; 0.105g/l potassium chloride; sodium bicarbonate 0.05g/l; and sodium chloride 0.05g/l) for
emulsification of oil sample. The diluted samples were then employed for subsequent analysis for microbial
content.
2.4 Microbiological Analysis of Sample
All the samples were analyzed for their viable bacterial and fungal load, fungi and bacteria that can
degrade palm oil, extracellular lipase producers as well as total coliform.
2.4.1 Enumeration of Total Heterotrophic Bacteria
The molten nutrient agar medium was employed and autoclaved at 121 oc for 15minutes.Oil samples
were serially diluted in the media in test tube before spreading onto sterile petri-dishes. One mililitre of the
dilutions 10-2,10-3 ,10-4.10-5,Were employed during plating. The agar medium was allowed to cool and solidify
before plates were incubated at room temperature for 24 48hrs. Colonies were picked from the plates to purify.
Pure culture of the bacteria were streaked on nutrient agar slant and stored in the refrigerator (4 0C) as stock
cultures for identification tests.
2.4.2 Enumeration of Total Heterotrophic Fungal
Potato Dextrose Agar (PDA) and Ringers solution was employed in this analysis. Rifampicin (an
antifungal agent) was added to PDA to suppress bacterial growth the agar medium was autoclaved at 121 0C for
15 minutes. Oil samples were serially diluted in molten agar in test-tubes before pouring into sterie petri dishes,
one milliliter of 10-5,10-7 dilution were used. The agar medium was allowed to cool and solidify before the plates
were incubated at room temperature (280c) for 72 120hrs.The counts for Fungal load was taken. A portion of
each fungal colony which developed was picked and sub-cultured onto Potato Dextrose Agar plates using
inoculating needle which was then kept as stock cultures for identification.
Enumeration of Total Coliform
Due to the immiscible nature of Palm kernel oil with an aqueous solution, solid Macconkkey medium
was employed for the enumeration of total coliforms. An aliquot from each prepared samples were plated onto
MacConkey agar medium and incubated at 370 C for 2 days
Isolation of Lipid Utilizing Bacteria (LUB)
Samples that have been serially diluted on a palm oil medium by a modification of the medium by
Sirisha et al., (2010) containing 0.5% (w/v) peptone, 0.3% (w/v) yeast extract, 1% (v/v) palm oil (sole carbon)
and 2% agar by spread plate method. Plates were incubated at 37 0C for two days. Pure cultures of the isolate
were maintained on nutrient agar (yeast extract) NaCl, peptone and 2% agar, pH 7.0) and were sub cultured
every 15 days.
Isolation of Lipid Utilizing Fungi (LUF)
Modified composed medium (15g peptone, 5g NaCl, Ig CaCl2, 10ml palm oil, 15g agar) was employed.
An aliqout of the sample was plated onto the medium which has palm oil as the sole source of carbon and
incubated for four days at 370C (Sirisha et al., 2010)
Screening Fungi for Lipase Activity Using The Rhodamin-B Agar Dye
Modified method by Savitha et al (2007). This method involves measurement of fluorescence caused
by the fatty acid released due to the action of lipase on olive oil. The quantitative fluorescence assay is based on
the interaction of Rhodamine B with fatty acid released during the enzyme hydrolysis of olive oil.
The fungi isolates were inoculated in media of the following composition (g/L). Potato dextrose agar, and
sodium chloride 4.0. The medium was adjusted to pH 7.0, autoclaved and cooled to 60 0C. Olive oil (13.25ml)
and 10ml of Rhodamin-B solution (0.001% w/v) was added with stirring and emulsified by mixing for 1 min.
The medium was allowed to stand for 10min at 600C to reduce foaming. 20ml of the medium was poured into
sterile petri dishes.
Screening Bacteria for Lipolytic Using The Rhodamin Bdye Agar
A solid medium composed of 0.5% (w/v) peptone, 0.3% (w/v) yeast extract, 1% olive oil and 10ml of
Rhodamin-B stock, 0.001% (w/v). The mixture was well homogenized by mixing. Cultures were incubated for
18-2hrs. For Rhodamin-B medium at 360C the lipolytic activities of isolates was monitored by fluorescence with

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Quality Assessment Of Unbranded Refined Palmkernel Oil In Distribution Within Five Local
UV light at 350nm. The isolates that produce extracellular lipase would hydrolyze the substrate and produce
halo zone surrounding the holes of the gar plates (Sirisha et al., 2010).
Identification of Isolates:
Identification of Bacterial Isolates
All bacterial isolates on the PCA plates were identified based on biochemical characteristics as
described by Cheesbrough, (2000) and U.S.FDA manual online (2001).
Identification of Fungal Isolates
All fungal isolates were identified based on their macroscopic and microscopic appearance with
reference to manual of Barnett and Hunter, (1972), Larone (1995) and Mycology online of Ellis (2006).
Physico-Chemical Analyysis of Samples:
Peroxide value, Iodine value, relative density Free-fatty acid and pH of each sample was analyzed
using methods employed by Pearson (1976) Ohimain et al., (2013) and Ohimain et al., (2012

Plate 1: Bacterial Colonies Under Uv Illumination. Orange Halo Is Due To Extracellular Lipase
Production By The Colony

Plate 2: Microscopic Characteristics Of Aspergillus Niger Isolated From Palm Kernel Oil From
Rumuokoro Market.

III.

Results And Discussion

The quality of palm kernel oil (PKO) depends on its physicochemical state and characteristics and also
on its microbiological quality. According to the stipulated standard by National Agency for Food and Drug
Administration and Control(NAFDAC).The acceptable limit of microorganism in palm oil and other vegetable
oils are 105ml for Aerobic mesophillic bacteria or total heterotrophic Bacteria.104ml for Aerobic mesophillic
fungus, 102ml for coliforms,( Okechalu et al.,2011 ). The differences in the results obtained for the total
heterotrophic bacteria and fungi load and the total lipid utilizing bacteria and fungi load may be due to the
presence of non lipid degrading microbes co-existing with lipid degraders for survival.
Table 1: Total heterotrophic bacterial and fungal count.
THBC
THFC
TOTAL
COLIFORMS

DOI: 10.9790/2380-08424550

OIL MILL
3.0 108
1.56106
1.6108

CHOBA
3.3108
2.2106
5.0108

MILE 1
1.51010
9.2105
3.0105

MILE 3
1.2108
1.4106
5.0107

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RUMUOKORO
1.2107
5.5105
4.2108

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Quality Assessment Of Unbranded Refined Palmkernel Oil In Distribution Within Five Local
Results obtained on the microbiological analysis (Table 1) carried out showed that the refined palm kernel oil
samples from all the five markets in Port Harcourt were above the microbiological quality standard of the
National Agency for food and drug administration and control.
Table 2: Lipid utilizing bacterial count
TOTAL LIPID
UTILIZING
BACTERIA
TOTAL LIPID
UTILIZING
FUNGI

OIL
MILL
MARKET
6.0107

CHOBA
MARKET
1.6108

MILE
MARKET
1.4109

2.0105

3.0105

7.0104

MILE 3 MARKET
3.3107

RUMUOKORO
MARKET
6.0106

6.0105

4.0104

The presence of coliform is an indicator of fecal contamination which could be as a result of the use of
untreated water or contaminated water and also as a result of bad handling during processing and packaging.
Growth on Rhodamine B agar and subsequent of exhibition of the characteristic yellow hyper orange
halo around the colony under ultra violet light signifies the secretion of extracellular lipase by the isolate. Upon
inoculation of isolates obtain from samples from market the presence of the orange halo which was expressed on
the Rhodamine B plate under the ultra violet light indicate the ability of the organisms to liberate extracellular
lipase. The orange halo on the Rhodamine plates (PLATE 1) shows the ability of the isolates to liberate
extracellular lipase which is known for its ability to degrade lipid liberating free fatty acids and glycerol.
Table 3: Physico-chemical characteristics of samples
SAMPLE
SITE
RUMUOK
ORO
CHOBA

IODINE
VALUE(wijis)
16.15

RELATIVE
DENSITY(g)
0.939

PEROXIDE
VALUE(Meq/kg)
59.85

FREE FATTY ACID(%)

pH

1.36

5.34

16.05

0.941

59.48

1.29

5.30

MILE 3
OIL MILL

17.81
15.87

0.949
0.905

59.55
59.93

1.57
1.41

5.38
5.32

MILE 1

15.93

0.918

59.76

1.35

5.37

The liberation of free fatty acids is an indication of spoilage. According to the food and Agricultural
organization (FAO) and world health organization (WHO) stipulate the acceptable free fatty acid value (Ffa) not
to exceed 1.376%. Most of these sample falls within the standard except for samples from Mile 3 and Mile 1
markets because of the high number of lipid utilizing bacteria isolated from the sample from these markets. The
low value obtained could suggest the onset of the breakdown of lipid molecules by exogenous lipases liberated
by lipolytic microorganisms. Acidic pH obtained may be due to the presence of liberated fatty acids.
Iodine number measures the degree of unsaturation of free fatty acids indicating that the higher the
degree the greater the likely-hood of rancidity. According to the FAO/WHO and CODEX standards, the limits
value for iodine number in grams is 50.0-55.0. The iodine value for these samples, where found to fall within
the limits (TABLE 3); an indication of the presence of saturated fat and a very low tendency to get rancid.
Peroxide value, reflects the state of oxidation and therefore the stability and quality of the oil. The
peroxide value of samples is found to exceed the stated standard this shows the high oxidation state of the
samples this could be the reason for the constant development of bad tastes in the palm kernel oil over a short
period of time. Peroxide value determines the degree of oxidation in oil as well as an indication of level of
deterioration of oil and fats( Nwanekezi and Onyeagba (2007); Okechalu (2011) . The high Peroxide value of
the samples indicates an onset of oxidation, which agrees with the reports by Ekpa and Ekpe (1996) who stated
that lipid degrading enzymes such as peroxidases and lipooxygenases ( Onyeka et al., 2005). Oxidation takes
place also, when microorganisms are capable of utilizing fatty acids in the absence of other simple sources of
carbon through a catalytic pathway known as -oxidation.
Relative density, is a physical measures of adulteration of vegetable oils, since different oils have
characteristic density and refractive index. Studies have shown that the contamination of vegetable oils with
particulate matters and other chemical adulterants such as potassium hydroxide brings chemical reaction with
fatty acids of vegetable oils with the production of soap i.e. carboxylic acid ester which alter the optical activity
of the vegetable oils and increases the susceptibility of the vegetable oils to become rancid or spoiled (Williams,
1990; Chabiri et al., 2009). The maximum acceptable limit of Relative density for palm kernel is within 0.8990.914 according to CODEX (1992).The results obtained on this were relative exceeding with slight differences.

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Quality Assessment Of Unbranded Refined Palmkernel Oil In Distribution Within Five Local

The study on the microbial profile of palm kernel oil carried out suggests the peculiarity of some
isolated sample from different markets Alcaligenes sp were found to be peculiar to Mile 1, Mile 3 and Oil mill
markets. Enterococcus, Pseudomonas and Staphylococcus species in all the markets which could be attributed
to poor handling and human contact.
Achromobacter sp were found in Mile 1 and Oil mill markets only, Micrococcus sp in Rumuokoro
markets where as Klebsiella sp was found in only Rumuokoro markets; in addition to bacteria, lipolytic fungi
such as Penicillium sp, Aspergillus niger and Fusarium species were found in all the samples from all the
markets.

IV.

Conclusion

The microbiological quality of samples which exceeded the standard quality suggests how poor the
traditional / local production process and poor handling of products have been. The presence of coliforms and
other indicates the possible presences of organism that could pose health hazards. But can be overcome over
boiling, interestingly, these products have been found to have the tendency to resist rancidity.
Therefore, more improvement is required in the production and distribution of groundnut oil in order to
meet the stipulated standards.

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